全 文 ：作物学报 ACTA AGRONOMICA SINICA 2012, 38(12): 2198−2205 http://www.chinacrops.org/zwxb/
ISSN 0496-3490; CODEN TSHPA9 E-mail: xbzw@chinajournal.net.cn

This work was supported by National Natural Science Foundation of China (30830074) and the project of the Department of Education of
Anhui Province (KJ2010B155).
Corresponding author: ZHANG Gui-Quan, E-mail: gzhang@scau.edu.cn
Received(收稿日期): 2012-04-09; Accepted(接受日期): 2012-07-05; Published online(网络出版日期): 2012-09-10.
URL: http://www.cnki.net/kcms/detail/11.1809.S.20120910.1331.008.html
DOI: 10.3724/SP.J.1006.2012.02198
Evaluation of Restorability of Two Fertility Restorer Genes in the Rice Chro-
mosome Single Segment Substitution Lines (SSSLs)
CAI Jian1 and ZHANG Gui-Quan2,*
1 School of Life Science, Fuyang Teachers College, Fuyang 236041, China; 2 State Key Laboratory for Conservation and Utilization of Subtropical
Agro-Bioresources, South China Agricultural University, Guangzhou 510642, China
Abstract: Rice chromosome single segment substitution line (SSSL) S15, a strong restorer line for WA-, and DA-CMS, was re-
cently isolated at Guangdong Provincial Key Laboratory of Plant Molecular Breeding, South China Agricultural University. The
present study was carried out with the objective to investigate the genetic mode of fertility restorer (Rf) genes and the genetic rela-
tionship between WA- and DA-CMS systems. The SSSL S15 was used to pollinate WA-CMS line of Bobai A and DA-CMS line
of Xieqingzao A for generation of two BC3F2 populations by using marker-assisted selection (MAS) and traditional backcrossing.
The results were as follows: (1) The genetic effect showed a trend of WA-CMS > DA-CMS in the genetic background of SSSL
S15, and the effect of Rf4 was slightly larger than that of Rf3 for the two CMS systems. (2) Two pairs of dominant genes governed
restoration of pollen fertility restoration, and some modifying or minor genes were involved in the inheritance of restorer ability
besides Rf3 or Rf4 in SSSL S15, indicating that the genetic mode of Rf genes showed a qualitative-quantitative characteristic for
WA-, and DA-CMS system. (3) When 202 SSR markers were used to analyze the genetic background and average length of sub-
stituted chromosome segments of the two BC3F2 individuals, carrying the genotypes Rf3Rf3/rf4rf4 or rf3rf3/Rf4Rf4, the mean of
segments of inherited background of them was 1.0, while the average lengths of the substituted chromosome segments, corre-
sponding to Rf3 and Rf4 loci, of them were 12.9 cM and 18.4 cM, respectively.
Keywords: Rice; Chromosome single segment substitution lines (SSSLs); Wild abortive (WA); Dwarf-wild-abortive (DA); Re-
storer gene (Rf )
基于水稻单片段代换系的 2个恢复基因恢复力的评价
蔡 健 1 张桂权 2,*
1 阜阳师范学院生命科学学院, 安徽阜阳 236041; 2 华南农业大学植物分子育种重点实验室, 广东广州 510642
摘 要: 由华南农业大学选育的水稻单片段代换系 S15对于野败型(WA)和矮败型(DA)细胞质雄性不育系均具有较强
的恢复性。以野败型不育系博白 A和矮败型不育系协青早 A为母本, 单片段代换系 S15为父本杂交, 采用分子标记
辅助选择和连续回交的方法构建了 2个 BC3F2群体。利用与第 1、第 10染色体上恢复基因 Rf3和 Rf4两侧紧密连锁
的 SSR 标记, 从这 2 个 BC3F2群体中筛选携带基因型 Rf3Rf3/rf4rf4 和 rf3rf3/Rf4Rf4 的单株, 观察这些单株花粉和小
穗育性, 并利用 202个多态性 SSR标记分析这些单株的遗传背景, 结果表明: (1)在同一细胞核背景下(S15), DA型细
胞质的可恢复性好于 WA 型细胞质, 单片段代换系 S15 中的恢复基因 Rf4 的恢复力大于恢复基因 Rf3 的恢复力。(2)
单片段代换系 S15 中的恢复基因对于 WA 型不育系博白 A 和 DA 型不育系协青早 A 表现出质量-数量性状的遗传。
在单片段代换系 S15 中, 除了主效恢复基因 Rf3 和 Rf4 外, 微效基因或者修饰基因也表现出对于博白 A 和协青早 A
的恢复性作用, 而且效应较大。(3)在构建的 2个 BC3F2群体中, 携带基因型 Rf3Rf3/rf4rf4和 rf3rf3/Rf4Rf4单株的遗传
背景片段数平均为 1.0, 对应于恢复基因 Rf3和 Rf4座位的代换片段平均长度分别为 12.9 cM和 18.4 cM。
关键词: 水稻; 染色体单片段代换系; 野败型(WA); 矮败型(DA); 恢复基因(Rf)
Cytoplasmic-nuclear male sterility (CMS) has been
found in over 150 plant species from 50 genera and 20
families [1-2], and is of significance in the production of
plant hybrids without requirement of manual emascula-
第 12期 蔡 健等: 基于水稻单片段代换系的 2个恢复基因恢复力的评价 2199


tion. CMS with maternal inheritance unable to make a
plant carrying mitochondrial defects produce functional
pollen, has been observed in numerous species[3]. In rice,
several CMS/Rf systems defined by different CMS cyto-
plasms with distinct genetic features have been identified.
These are wild abortive (WA), dwarf-wild-abortive (DA),
boro type II (BT), and Honglian (HL) CMS systems. WA
(indica) and DA (indica), discovered in China, belong to
the sporophytic CMS system and both of them possess
typical aborted pollens, whereas HL (indica) and BT
(japonica) CMS systems are categorized as gametophytic
types [4-7].
CMS can be restored by fertility restorer (Rf) genes
associated with nuclear genes encoding pentatricopeptide
repeat (PPR) proteins [8]. Although fertility can be re-
stored by the dominant restorer genes, reports on the
number, positions, and the effects of these Rf genes are
inconsistent. Currently, there are over 17 Rf alleles for
different CMS systems were reported in rice. Apart from
Rf2, Rf3, and Rf17 mapped on chromosomes 2, 1, and 4,
respectively [9-11], others including Rf4 for the WA-CMS
system, Rf1 for BT-CMS, Rf5 and Rf6 for HL-CMS, and
qRf-10-2 for DA-CMS, etc., reside on the long arm of
chromosome 10, and are closely linked to form a gene
cluster[5,12-19].
To better understand the relationship and the genetic
effect of Rf genes of different cytoplasmic sources and
the restoring abilities of Rf genes in wild and cultivated
rice for WA- and DA-type CMS, we have constructed a
library of 1 123 single-segment substitution lines (SSSLs)
in rice [20], and has used it to detect QTLs affecting many
agronomic traits in rice [21-22]. SSSLs are the powerful
tools to dissect complex traits into a set of monogenic
loci, to assign phenotypic values to different alleles at the
loci of fertility restorer (Rf) genes and to study the rela-
tionship between Rf and CMS [20,23]. However, no re-
search has so far been conducted to study the genetic
relationship among WA and DA CMS systems, and the
genetic mode of Rf genes from different cytoplasmic
sources utilizing the library of SSSLs. This study aimed
at (1) studying genetic mode of Rf genes for WA- and
DA-CMS systems, and (2) evaluating the genetic
relationship between WA- and DA-CMS systems, and (3)
studying the effect of Rf genes on fertility restoration of
individual plants in BC3F2 populations.
1 Materials and Methods
1.1 Plant materials
The SSSLs in the library were developed by using of
Huajingxian 74 (HJX74), an elite indica variety from
Southern China, as a recipient, and 24 varieties including
14 indica and 10 japonica varieties collected worldwide
as the donors [20]. The development of the SSSLs,
through backcrossing and SSR marker selection, was
described by He et al. [21] and Xi et al. [22] For this study,
an SSSL i.e. W20-06-06-11 (code S15), with the geno-
type Rf3Rf3/Rf4Rf4, a strong restorer line for WA- and
DA-CMS, was recently identified by State Key Labora-
tory for Conservation and Utilization of Subtropical
Agro-Bioresources, South China Agricultural University.
Gene Rf3 on chromosomes 1 was introgressed from the
recipient HJX74, and Rf4 on chromosomes 10 from the
donor Chenlongshuijing (an indica variety from China).
A typical wild-abortive CMS lines of Bobai A (BbA,
indica) and a typical dwarf-wild-abortive CMS line of
XieqingzaoA (XqA, indica) were used as the female par-
ents (A-lines) to cross with SSSL S15. These plant
materials and their derived progenies were all planted in
the experimental field in South China Agricultural Uni-
versity campus during the 2007–2009 year.
1.2 Population construction
Two BC3F2 populations, possessing the same genetic
background of SSSL S15 and different CMS, such as
WA- and DA-types, were constructed by using marker-
assisted selection (MAS) and backcrossing SSSL S15 as
the male parent to BbA and XqA for four times, respec-
tively, to determine the genetic mode of Rf3 and Rf4
based on the marker genotypes. Eight hundreds fifteen
simple sequence repeat (SSR) markers, distributed across
12 chromosomes, selected on the rice microsatellite
maps[24] and designed by this study, were used to survey
the polymorphism between each CMS line and HJX74.
The polymorphism in the two crosses was 25.2% and
24.4%, respectively, with a mean of 24.8%. The poly-
morphic markers were used for MAS in the process of
BC3F2 population development, which consisted of se-
lecting genetic background and target genes Rf3 and Rf4
of individuals from F1 to BC3F2 progenies. The average
size of the intervals between polymorphic markers in the
two crosses was 7.3 cM and 7.5 cM with a mean of 7.4
(Table 1). In the two BC3F2 populations, plants, carrying
genotypes Rf3Rf3/rf4rf4 and rf3rf3/ Rf4Rf4 were deter-
mined using the corresponding SSR markers closely
linked to Rf3 and Rf4 genes (Table 2). SSSL S15, two
CMS lines and their derived progenies were all planted in
the experimental field within South China Agricultural
University campus during the 2007–2010 year.
1.3 SSR marker analysis
Mini-scale DNA extraction was carried out according
to the procedure described by Zheng et al. [25] The PCR
was conducted according to Panaud et al. [26] with minor
modifications. The PCR program was as follows: 94°C
for 5 min, followed by 35 cycles of 94°C for 1 min, 55°C
for 1 min, 72°C for 1 min, and a final extension at 72°C
for 5 min. The PCR products were separated through
electrophoresis on 6% polyacrylamide gels. Bands were
visualized following a silver staining.
1.4 Estimation of number and length of substi-
tuted chromosome segments
In the two BC3F2 populations, the number of genetic
2200 作 物 学 报 第 38卷

Table 1 Polymorphic SSR markers between HJX74 and A-lines
A-lines
Description of markers
BbA XqA
Average
Total number of markers tested 815 815 815
No. of polymorphic markers 205 199 202
Ratio of polymorphism of markers (%) 25.2 24.4 24.8
Average size of the interval between polymorphic markers (cM) 7.3 7.5 7.4
Distribution of polymorphic markers on 12 chromosomes
Chr.1 29 28 28.5
Chr.2 18 18 18.0
Chr.3 21 20 20.5
Chr.4 16 12 14.0
Chr.5 14 14 14.0
Chr.6 13 14 13.5
Chr.7 13 15 14.0
Chr.8 15 13 14.0
Chr.9 15 17 16.0
Chr.10 21 13 17.0
Chr.11 13 15 14.0
Chr.12 17 20 18.5


Table 2 Primer sequences of the SSR markers used for detecting Rf3 and Rf4 genes in this study
Code Marker Linked Rf gene Chr. Primer sequence (5–3) Reference
1 RM1 Rf3 1 F: gagaaaacacaatgcaaaaa; R: gcgttggttggacctgac McCouch et al. [24]
2 PSM348 Unreported F: gatgaggttaggttggtgcc; R: gtagaatcaactcgagcggc Not reported
3 PSM354 Unreported F: acaagctaaggtagtgtccatg; R: cattttacctcaggctcttca Not reported
4 PSM572 Unreported F: aagttgctcgggactaaaat; R: caggtaataagggtagaggg Not reported
5 RM304 Rf4 10 F: gatagggagctgaaggagatg; R: tcaaaccggcacatataagac McCouch et al. [24]
6 RM5373 Rf4 F: attgctccttacctccttgc; R: ggagatgctatagcagcagtg McCouch et al. [24]
7 RM6100 Rf4 F: tcctctaccagtaccgcacc; R: gctggatcacagatcattgc McCouch et al. [24]
8 PSM25599 Unreported F: cctgcagtactcgcggaagagg; R: ggacgaacaccagtaggatctcagg Not reported


background segments and the length of substituted chro-
mosome segments with Rf loci in the plants with respec-
tive genotypes, such as Rf3Rf3/rf4rf4 and rf3rf3/Rf4Rf4,
were estimated based on graphical genotypes [27]. A chro-
mosome segment flanked by two markers of donor type
(DD) is considered as the donor genome, a chromosome
segment flanked by two markers of recipient type (RR) is
considered as the recipient genome, and a chromosome
segment flanked by one marker of donor type and one
marker of recipient type (DR) is considered as 50% donor
and 50% recipient genome. In other words, the length of
DD plus the length of two half DR was considered to be
the estimated length of a substituted chromosome segment.
1.5 Determination of pollen
For all individuals selected in the two BC3F2 of
(A-lines)/SSSL S15, pollen fertility was investigated.
Five anthers were taken randomly from the spikelets col-
lected to identify pollen fertility. Pollen fertility was used
as the main criteria for the evaluation of fertile and sterile
plants. Mature anthers were harvested, and the pollen
was stained with 1 % I2-KI solution. The numbers of dark
blue (stainable) and clear pollen grains (unstainable) in
each individual were counted under an optical micro-
scope described by Zhang and Lu [28].
2 Results
2.1 Genetic relationship among WA and DA
CMS systems
Two BC3F2 populations in which plants possessed the
same nuclear genetic background of SSSL S15 and di-
verse CMS, such as WA- and DA-types, were developed
by backcrossing SSSL S15 to two alloplasmic CMS lines
(A-lines) of the respective cytoplasm for four times, in-
volving BbA and XqA, respectively, and selecting the
第 12期 蔡 健等: 基于水稻单片段代换系的 2个恢复基因恢复力的评价 2201


seedlings that showed the maximum number of SSSL
S15-type bands using SSR markers covering the whole
genome. The BC1F1 plants of BbA/S15 and XqA/S15,
which were exhibited the genotypes Rf3rf3/Rf4rf4, were
screened for the markers that were closely linked to two
Rf loci on chromosomes 1 (Rf3) and 10 (Rf4). Then, a
genome-wide scan was carried out to determine the
maximum number of SSSL S15-type bands using 205
and 199 SSR markers in the plants (Table 1), homozy-
gous or heterozygous at the locus, of BbA/S15 and
XqA/S15, respectively. The selected plants were back-
crossed to SSSL S15 and re-screened for the markers
linked to Rf3 or Rf4 locus, followed by the SSR markers
for three generations, to produce two BC3F2 populations.
In the two BC3F2 populations, the plants, carrying the
genotypes Rf3Rf3/rf4rf4 and rf3rf3/Rf4Rf4 were detected,
respectively, and their phenotyping for pollen and
spikelet fertility were evaluated (Table 3). The results
showed that across the two crosses involving A-lines and
SSSL S15, the spikelet fertility showed a trend of XqA/
S15 (89.64%) > BbA/S15 (70.76%), the pollen fertility
showed a trend of XqA/S15 (51.71%) > BbA/S15
(47.82%), indicating that the genetic effect of two CMS
would be WA-CMS > DA-CMS in the genetic back-
ground of SSSL S15 in rice.
2.2 Genetic effects of two Rf loci
The two Rf loci acted as a pair of classical dominant
genes in two BC3F2 individuals derived from the crosses
involving A-lines and SSSL S15 (Table 4). In the two
BC3F2 populations, for the plants carrying the genotype
Rf3Rf3/rf4rf4, the average values of phenotyping for
pollen (spikelet) fertility were 45.81% (78.77%), with a
mean of 62.29%. For the plants of the two BC3F2 popula-
tions, which carried genotypes rf3rf3/Rf4Rf4, the average
values of phenotyping for pollen (spikelet) fertility were
53.71% (81.63%), with a mean of 67.67%. Therefore, the
increasing order of pollen and spikelet fertility for WA-
and DA-CMS would be rf3rf3/Rf4Rf4 > Rf3Rf3/rf4rf4,
which indicated that the effect of Rf4 appeared to be
slightly larger than that of Rf3.

Table 3 Pollen (spikelet) fertility (%) of BC3F2 individuals of A-lines/S15 cross-combination for WA and DA CMS (%)
Cross Genotype Pollen fertility a Range Spikelet fertility a Range Average
BbA/S15 Rf3Rf3/rf4rf4 43.41±0.24 31.42–68.12 69.32±0.35 30.59–90.71
rf3rf3/Rf4Rf4 52.22±0.17 41.31–81.21 72.20±0.60 40.60–93.44
Average 47.82 70.76 59.29
XqA/S15 Rf3Rf3/rf4rf4 48.21±0.31 17.31–72.10 88.21±1.32 45.16–94.12
rf3rf3/Rf4Rf4 55.20±0.45 22.32–83.21 91.06±1.81 50.71–96.85
Average 51.71 89.64 70.67
a Mean ± SE.

Table 4 Comparison of restoring ability for two restorer genes (Rf3 and Rf4) with two CMS lines in the two BC3F2 populations (%)
Cross Genotype Pollen fertility a Spikelet fertility a Average
BbA/S15 Rf3Rf3/rf4rf4 43.41±0.24 69.32±0.35
XqA/S15 48.21±0.31 88.21±1.32
Average 45.81 78.77 62.29
BbA/S15 rf3rf3/Rf4Rf4 52.22±0.17 72.20±0.60
XqA/S15 55.20±0.45 91.06±1.81
Average 53.71 81.63 67.67
a Mean ± SE.

2.3 Genetic analysis of fertility restorer genes in
SSSL S15
In the two BC3F2 populations, derived from the cross-
A-lines/S15, the plants, carrying the genotypes Rf3Rf3/
rf4rf4 and rf3rf3/Rf4Rf4, were detected, respectively, and
their phenotyping for pollen fertility was evaluated to
understand the genetic patterns of the fertility restorer
genes for the two CMS lines. The distribution of pollen
fertility of the BC3F2 plants is shown in Fig. 1. It could
be concluded from Fig. 1 that pollen fertility of the
BC3F2 plants showed something like a continuous single
peak distribution, indicating that restorer ability of SSSL
S15 was controlled by a dominant gene with some
modifying or minor genes. The pollen fertility of the
BC3F2 plants (BbA/S15) with the genotypes Rf3Rf3/
rf4rf4 and rf3rf3/ Rf4Rf4 ranged from 31.42% to 68.12%
and 41.31% to 81.21%, respectively (Fig. 1-A and B),
while the pollen fertility of BC3F2 plants (XqA/S15) with
the genotypes Rf3Rf3/rf4rf4 and rf3rf3/Rf4Rf4 ranged
from 17.31% to 72.10% and 22.32% to 83.21%, respec-
tively (Fig. 1-C and D), which indicated that the effect of
Rf4 appeared to be slightly larger than that of Rf3. So, for
2202 作 物 学 报 第 38卷

WA-, and DA-CMS systems, the genetic mode of ferti-
lity restorer (Rf) genes showed a qualitative-quantitative
characteristic governed by two pairs of dominant genes
and some modifying or minor genes in SSSL S15.

Fig. 1 Distribution of pollen fertility of the BC3F2 plants carrying the genotypes Rf3Rf3/rf4rf4 and rf3rf3/Rf4Rf4, derived from the
cross-A-lines/S15
A and B indicate the plants (BbA/S15) with the genotypes Rf3Rf3/rf4rf4 and rf3rf3/Rf4Rf4, respectively, and C and D indicate the plants (XqA/S15)
with the genotypes Rf3Rf3/rf4rf4 and rf3rf3/Rf4Rf4, respectively.

2.4 Number and length of substituted chromo-
some segments of individual plants in BC3F2 popu-
lations
Using SSR markers (Table 1 and Table 2), a set of
BC3F2 plants with the genotypes Rf3Rf3/rf4rf4 and
rf3rf3/Rf4Rf4 were identified from the two BC3F2 popu-
lations, derived from the cross-A-lines/S15, and then a
genome-wide scan was carried out to determine the ho-
mozygous or heterozygous at the locus in the plants. The
number of donor segments of the individual plants in the
BC3F2 population (BbA/S15) was 0.8 and 1.1, respec-
tively (Table 5), whereas it was 1.2 and 0.9 in the BC3F2
population (XqA/S15), respectively. The length of the
substituted chromosome segments corresponding to Rf3
and Rf4 locus of the BC3F2 plants (BbA/S15) was 15.8
cM and 16.8 cM, while it was 9.9 cM and 20.0 cM of
BC3F2 plants (XqA/S15), respectively.
3 Discussion
The numbers and acting model of Rf genes are specific
to each of the CMS types within a plant species [29].
Some references have suggested that different Rf alleles
interact with CMS in a gene-for-gene fashion, this means
that various Rf loci are correspondingly determined by
the multiple CMS systems existed in the natural popula-
tions within a plant species [30-31]. In this research, the
increasing order of genetic effect of the two CMS would
be WA-CMS > DA-CMS in the genetic background of
SSSL S15 in rice (Table 3), similar to the results obtained
by Fu et al. [32] and Xie et al. [5], who reported that the
effect of WA-CMS appeared to be larger than that of
DA-CMS. For the WA- and DA-CMS systems, the effect
of Rf4 appeared to be slightly larger than that of Rf3 (Ta-
ble 4), similar to the results reported previously [12, 33-35].
Differences in degree of fertility restoration in differ-
ent cytoplasmic backgrounds had been reported by Prad-
han and Jachuck [36], who attributed such differences to
the influence of the female genotype and/or differential
penetrance of restorer genes in different CMS lines. Cer-
tain modifier genes could also be responsible for differ-
ences in fertility levels. In this paper, Fig. 1 shows that
the genetic mode of fertility restorer (Rf) genes is a
qualitative-quantitative trait governed by two pairs of
dominant genes and some modifying or minor genes in
SSSL S15. Similar results were obtained by Zhuang et
al. [34] and Tao et al. [37], who reported that the restoring
genes in WAB450-11 and IRAT359 were non·allelic to
Rf1 in C57 or some modifying or minor genes were in-
volved in sterility restoration besides Rf1. Here, we ana-
lyzed the genetic relationship among WA- and DA-CMS
in the same nuclear genetic background of SSSL S15,
第 12期 蔡 健等: 基于水稻单片段代换系的 2个恢复基因恢复力的评价 2203


Table 5 Substituted chromosome segments in the BC3F2 plants (A-lines/S15) carrying the genotypes Rf3Rf3/rf4rf4 or rf3rf3/Rf4Rf4
Position 1)
Cross No. of plants Genotype
No. of genetic
background
segment P1 P2 P3 P4
Marker 2) Length(cM)
Substituted chromosome segments corresponding to Rf3 locus
BbA/S15 55 Rf3Rf3/rf4rf4 0.8 23.3 24.7 30.5 49.0 RM220--RM1-RM283-PSM572- PSM348-PSM354-RM259--RM581 15.8
57 rf3rf3/Rf4Rf4 1.1
XqA/S15 40 Rf3Rf3/rf4rf4 1.2 23.3 24.7 28.9 38.8 RM220--RM1-RM283-PSM347-RM151- PSM572-PSM348-PSM354--RM259 9.9
37 rf3rf3/Rf4Rf4 0.9
Average 47.3 1.0 23.3 24.7 29.7 43.9 12.9
Substituted chromosome segments corresponding to Rf4 locus
BbA/S15 55 Rf3Rf3/rf4rf4 0.8
57 rf3rf3/Rf4Rf4 1.1 48.8 49.0 61.0 70.0
RM258--PSM25599-PSM25510-
RM304-RM5373-RM6100-PSM168-
RM271-PSM169--RM147
16.8
XqA/S15 40 Rf3Rf3/rf4rf4 1.2
37 rf3rf3/Rf4Rf4 0.9 32.0 49.0 59.0 61.0
PSM166--RM258-PSM25599-
RM304-RM5373-RM6100-
PSM168-RM271--PSM169
20.0
Average 47.3 1.0 40.4 49.0 60.0 65.5 18.4
1) P indicates the position of substituted segment on chromosome 1 or 10. P1 and P4 indicate the position of maximum length of substituted
segment, and P2 and P3 indicate the positions of minimum length of substituted segment. The single hyphen in the middle of markers indicates chro-
mosome substitution segments. The end markers of the triadic hyphens are side markers of substitution segment which indicates that segment recom-
bination might appear.
2) RM code indicates the markers described in McCouch et al. [24], and PSM code indicates the markers designed by Guangdong Provincial Key
Laboratory of Plant Molecular Breeding, South China Agricultural University, China.

which not only avoided the interference of the nuclear
genetic background, but also provided the precise and
reliable results.
4 Conclusion
SSSL S15, a strong restorer line for WA- and DA-
CMS, was recently isolated from the library of SSSL in
rice. We can transfer the Rf genes in SSSL S15 into
adapted cultivars through a backcrossing program in an
active hybrid rice breeding program.
The genetic effect showed a trend of WA-CMS >
DA-CMS in the genetic background of SSSL S15
(HJX74) in rice, and the effect of Rf4 appeared to be
slightly larger than that of Rf3 for the two CMS systems.
For WA- and DA-CMS, the genetic mode of fertility
restorer (Rf) genes showed a qualitative-quantitative
characteristic governed by two pairs of dominant genes
and some modifying or minor genes in SSSL S15.
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